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一个新的东亚钳蝎毒素(BmKT1)全长cDNA的克隆和分析

Cloning and Characterization of the Full Length cDNA Sequence of a New Venom Peptide(BmKT1)from Chinese Scorpion Buthus martensii Karsch

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【作者】 曾宪春李文鑫朱智慧朱顺义彭方毛歆

【Author】 ZENG Xian chun,LI Wen xin ** ,ZHU Zhi hui,ZHU Shun yi,PENG Fang,MAO Xin (Department of Biotechnology,School of Life Sciences,Wuhan University,Wuhan 430072,China)

【机构】 武汉大学生命科学学院生物技术系!武汉430072

【摘要】 首先构建了东亚钳蝎毒腺组织 c DNA文库 ;根据已知的东亚钳蝎哺乳动物毒素氨基酸序列保守区设计引物 ,并用 PCR从 c DNA文库中扩增出一个 c DNA片段作为筛选 c DNA文库的探针 ;从 c DNA文库中筛选到二个编码同一个新的蝎毒素多肽的 c DNA,它们除 3′- UTR外 ,其余序列完全一致 .它们均含有 2 55bp长的开放阅读框 ,编码 85肽的前体毒素 ,包括 1 9个氨基酸残基的信号肽 ,66个残基的成熟毒素 (命名为 Bm KT1) ;Bm KT1氨基酸序列与已知的蝎毒素具有较大的同源性 ,与 Bm KM1,Lqq ,Lqhα IT和 Bm K M10 的同源性分别为 77%、67%、67%和 65% .Bm KT1的 C端不存在末端修饰步骤且具有一个与这些毒素不相同的特征结构 ,即在末端延伸了两个氨基酸残基 - P- S,推测 Bm KT1具有新的活性功能特征 .

【Abstract】 A cDNA library was constructed from the venom gland of Chinese scorpion Buthus martensii Karsch(BmK).A cDNA probe for screening the cDNA library was obtained by PCR amplification with primers designed and synthesized according to the conserved sequences of BmK mammalian neurotoxins.With the probe,two cDNAs of a scorpion toxin were isolated from the library.Except for their 3′ UTR,other sequences of the two cDNAs were identical.Both the cDNAs displayed an ORF(open reading frame)of 255 bp,which encoded a toxin precursor of 85 residues including a signal peptide of 19 residues and a mature peptide(named BmKT 1)of 66 residues.The analysis of amino acid sequence similarity indicated that the BmKT 1 was homologous with some scorpion toxins,showing 77%,67%,67%,65% sequence identity with that of BmKM 1,LqqⅢ,Lqh α IT,BmKM 10 respectively.The C terminus of BmKT 1 has no modification and has a peculiar end structure compared with other toxins,showing two extended residues P S.The difference in 3′ UTR between the two cDNAs suggested that they were evolved from a common ancestor gene and an interesting evolution mechanism was underwent by the scorpion for adjusting to various environment.

【关键词】 东亚钳蝎(Buthus martensii Karsch)cDNA毒素
【Key words】 Buthus martensii KarschcDNAToxin
【基金】 国家医药技术创新博士基金!( 96-90 1-16-0 3 3 );武汉市青年科技晨光计划!( 965 0 0 10 3 7-2 4);国家自然科学基金!( 3 9970 897)
  • 【文献出处】 中国生物化学与分子生物学报 ,CHINESE JOURNAL OF BIOCHEMISTRY AND MOLECULAR BIOLOGY , 编辑部邮箱 ,2000年05期
  • 【分类号】Q78
  • 【被引频次】7
  • 【下载频次】113
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